Methods for preparation of macrocyclic molecules macrocyclic molecules prepared thereby and substrates and solid supports for use therein

A solid support of claim 4, wherein

• m and n are independently selected integers from about 1 to about 6;
• m+n is from about 4 to about 8;
• E is O;
• and
• X is —S(O)— or —S(O)2—.

A solid support according to Formula IV:

• 34
• wherein
• E is O or S;
• t is an integer from about 1 to about 9;
• and
• Bead is a solid particle having amino functional groups to which a linker can be attached.

A method of preparation of a solid support of Formula I, comprising:

• providing a solid particle having a plurality of amino functional groups;
• contacting the bead with a carboxylic acid of the formula:
• 35
• E is S or O;
• p is an integer from 0-2;
• and
• Linker comprises a linear backbone comprising between about 4 carbon atoms and about 20 carbon atoms and between about 0 and 10 hetero atoms selected from N, O or S, where each carbon of the linear backbone may be optionally substituted with 0, 1, or 2 groups selected from C1-6alkyl, hydroxy, amino, halogen, C1-6alkoxy, or oxo;
• under conditions conducive to the formation of a solid support according to Formula I:

A method of preparation of a solid support of Formula I where p is 1, comprising:

• providing an insoluble polymer having a plurality of amino functional groups;
• contacting the polymer with a carboxylic acid of the formula:
• 36
• E is S or O;
• R is C1-6alkyl;
• Linker comprises a linear backbone comprising between 2 and 12 carbon atoms and from 0 to 6 heteroatoms selected from N, O and S in the linear backbone, where each carbon of the linear backbone may be optionally substituted with 0,1, or 2 groups selected from hydrogen, C1-6alkyl, hydroxy, amino, halogen, C1-6alkoxy, or oxo;
• under conditions conducive to the formation of a functionalized solid support according to the formula:
• 37
• contacting the functionalized solid support with β-alanine-O-methyl ester under conditions conducive to coupling the formation of an amide bond;
• contacting the polymer having a β-alaninyl coupled thereto with ethanolamine under conditions conducive to the formation of a solid support according to Formula II.

A method of formation of a library of macrocyclic molecules comprising:

• providing a plurality of solid support bound substrates suitable for macrocyclization according to Formula V:
• 56
• A is a peptidic sequence, synthetic hydrocarbon group, a polyketide, —(CH2CH2O)s—, or a combination thereof having at least about 10 atoms in a linear backbone;
• s is an integer from 1 to about 10;
• Nuc is NH2 or OH;
• BeadE is a solid support having at least one amino functional group;
• where each solid support substrate of the library of solid support bound substrates comprises a chemically distinct combination of A and Nuc;
• and
• contacting purified excised TE domain protein with each solid support bound substrate having a chemically distinct combination of A and Nuc under conditions conducive to formation of an transient TE-O-acyl bond and subsequent formation of a macrocyclic product by displacement of a TE domain by a pendant nucleophile, Nuc such that a plurality of chemically distinct macrocycles are formed.

A macrocycle according to Formula XIII:

• 59
• wherein
• B is a polyketide residue having between about 2 and about 12 carbon atoms in a linear chain which may include one or more amide linkages or double bonds in the linear chain and which may be optionally substituted with between one and 12 groups selected from C1-6alkyl, C2-6alkenyl, C2-6alkynyl, C3-8cycloalkyl, hydroxy, or C1-6alkoxy;
• R, Ri, and Rj are independently selected for each value of i from 1 to a and for each value of j from 1 to b from the group consisting of hydrogen, synthetic and biosynthetic amino acid residue side chains including side chains selected from C1-6alkyl, C2-6alkenyl, C2-6alkynyl, C3-8cycloalkyl, C3-8cycloalkylC1-4alkyl, hydroxy, C1-6alkoxy, hydroxyC1-6alkyl, thioC1-6alkyl, amino, mono and di(C1-6alkyl)amino, saturated, partially unsaturated or aromatic heterocyclic groups having 5 to about 15 ring atoms, 1 to about 3 rings and 1, 2, or 3 N, O or S ring atoms, aryl, arylmethyl, and heteroarylmethyl, where
• each amino acid residue side chain may be optionally substituted with one or more substituents selected from the group consisting of hydrogen, chloro, fluoro, bromo, iodo, cyano, nitro, amino, hydroxy, C1-6alkyl, aryl, benzyl, carboxylate, carbamoyl, thiol, C1-6alkylthiol;
• a and b are independently selected integers from about 2 to about 10;
• and
• X is O or NH.